CRISPR editing of sftb-1/SF3B1 in Caenorhabditis elegans allows the identification of synthetic interactions with cancer-related mutations and the chemical inhibition of splicing


Autoři: Xènia Serrat aff001;  Dmytro Kukhtar aff001;  Eric Cornes aff002;  Anna Esteve-Codina aff003;  Helena Benlloch aff001;  Germano Cecere aff002;  Julián Cerón aff001
Působiště autorů: Modeling human disease in . Group, Genes, Disease and Therapy Program, Institut d’Investigació Biomèdica de Bellvitge–IDIBELL, Barcelona, Spain aff001;  Mechanisms of Epigenetic Inheritance, Department of Developmental and Stem Cell Biology, Institut Pasteur, UMR3738, CNRS, Paris, France aff002;  CNAG‐CRG, Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, Barcelona, Spain aff003;  Universitat Pompeu Fabra (UPF), Barcelona, Spain aff004
Vyšlo v časopise: CRISPR editing of sftb-1/SF3B1 in Caenorhabditis elegans allows the identification of synthetic interactions with cancer-related mutations and the chemical inhibition of splicing. PLoS Genet 15(10): e32767. doi:10.1371/journal.pgen.1008464
Kategorie: Research Article
doi: 10.1371/journal.pgen.1008464

Souhrn

SF3B1 is the most frequently mutated splicing factor in cancer. Mutations in SF3B1 likely confer clonal advantages to cancer cells but they may also confer vulnerabilities that can be therapeutically targeted. SF3B1 cancer mutations can be maintained in homozygosis in C. elegans, allowing synthetic lethal screens with a homogeneous population of animals. These mutations cause alternative splicing (AS) defects in C. elegans, as it occurs in SF3B1-mutated human cells. In a screen, we identified RNAi of U2 snRNP components that cause synthetic lethality with sftb-1/SF3B1 mutations. We also detected synthetic interactions between sftb-1 mutants and cancer-related mutations in uaf-2/U2AF1 or rsp-4/SRSF2, demonstrating that this model can identify interactions between mutations that are mutually exclusive in human tumors. Finally, we have edited an SFTB-1 domain to sensitize C. elegans to the splicing modulators pladienolide B or herboxidiene. Thus, we have established a multicellular model for SF3B1 mutations amenable for high-throughput genetic and chemical screens.

Klíčová slova:

Alleles – Caenorhabditis elegans – CRISPR – Larvae – Mutant strains – RNA interference – Sequence motif analysis – Missense mutation


Zdroje

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